GSM7178923: H3K36me2_sh4_Input; Homo sapiens; ChIP-Seq
Tissue or cells are cross-linked with formaldehyde to form covalent bonds between proteins and DNA. The cells are then lysed to releasecytoplasmic contentsand collect the nuclei. DNA-protein complexesare extracted from the lysate, and the DNA is fragmented into 200-1000bp pieces. The target protein-DNA complex is immunoprecipitated and washed. The complex is then eluted, and the DNA-protein cross-links are reversed, and DNA is extracted. The quality of theprecipitated DNAis checked, and the protein is detected using Western blotting. Finally, theimmunoprecipitated DNAis used to construct a library. DNA from the IP was end-repaired and had an A-tail added. Sequencing adapters were then added to the repaired DNA. Target-sized fragments were recovered using magnetic beads and PCR amplified to obtain a library for sequencing. The library was quality checked using gel electrophoresis and quantified using Qubit 2.0 to ensure proper concentration for sequencing. Once the library passed quality control, it was sequenced on an Illumina sequencer, with different libraries sequenced based on their effective concentration and desired data output.